The rapidly changing field of flow cytometry has required the CCR-Frederick Flow Cytometry Core to replace old instrumentation and upgrade the newer instruments that we have to meet the requirements of CCR investigators. We purchased 3 new instruments in FY09. Our new LSRII detects 16 colors, with 3 lasers and digital data capabilities that will soon be enhanced with a green laser to enable the analysis of the new red fluorescent reporter proteins. The FACSAria SORP will also be equipped with a new green laser so that cells expressing these new fluorescent proteins can be sorted for further experiments. These experiments will allow several genes and their interactions with each other to be tracked using the new fluorescent reporter proteins. The green laser excites phycoerythrin (a red protein from the light-harvesting phycobiliprotein family) more efficiently, which will be useful as a very bright fluorchrome to detect dimly expressed epitopes such as the Kir protein. The green lasers were acquired at the request of several CCR laboratories to serve projects that these labs are planning to conduct in the near future. Both FACSAria sorters will have fluidics upgrades that will convert these instruments into the new model FACSAria II. The old BD FACSort will be replaced with a new FACSCalibur and the old LSRI will be replaced with a new 2-laser LSRII Fortessa. All of this new technology will be installed by the end of this year. The acquisition of these new instruments required more space with a new configuration to fit the needs of the Flow Cytometry Core facility. The new space was designed, renovated, and the work coordinated so that at each step instrument downtime was minimized to just a few hours. Flow Cytometry Core staff members were involved in the design and analysis of a Cancer and Inflammation Program (CIP)-wide collaboration to characterize the inflammatory response of tumor-infiltrating lymphocytes from 5 mouse tumor models using a common protocol to disaggregate the tumors and a common antibody panel so that the results would be standardized. These tumors have very different origins and disease characteristics because they are derived from mouse lung, breast, prostate, renal, and melanoma cancers. This data has provided information on the diverse micro-environments of each of these cancers, focusing on the inflammatory response of the host. During the first 9 months of this year, the Flow Cytometry Core staff trained more than 20 investigators to run their own samples. Staff members taught the standard flow cytometry technology to novice users. More experienced investigators, who require complex experiments to answer their research questions, were trained on the facility's newer digital instruments, which are capable of sorting more antibody combinations. Flow Cytometry Core staff members operate all of the cell sorters due to the complexity of these instruments. During the last year, we sorted more than 500 samples. We also analyzed more than 9000 samples for those investigators who use this technology less often (or those who are not trained to run the analyzers) and for those investigators who prefer to have the Core staff conduct the analysis. The productivity of the Flow Cytometry Core has been greatly increased by training individual investigators to analyze their own samples. The instruments are used daily, often into the night and on the weekends. In the past year, data has been generated (or samples have been sorted) for 102 individual scientists from the laboratories of 37 principal investigators. Although the CCR-Frederick Flow Cytometry Core is part of the CIP, investigators from this program account for approximately 40% of the use of this facility.